As analytical methods for the secretion ratio of similar structural proteins in a test sample, enzyme immunoassay (EIA) and radioimmunoassay (RIA) etc. are currently used for quantitative determination of individual analyte, and the relative ratio is measured and used for a diagnosis of disease. However, because the methods as mentioned above carry out the analysis of analytes by using monoclonal antibody which recognizes specific epitope, the analytical result differs respectively according to analytical method, further shows a different numerical value in accordance with standard. As such, determination of the ratio based on analytical values showing difference even in a test sample of same patient naturally includes a large error, so it is hard to be used as objective clinical result.
It was known that polycystic ovary syndrome (PCOS) shows a high incidence of about 5% in teenagers and adults, respectively, and relatively common in fertile young women. The size of ovary in PCOS patients is 2-3 times larger than that of normal women, and about 10 small sacs are found therein. It has been reported that even in case of normal women, about 30% thereof shows polycystic ovary symptom on ultrasonography, yet in case of infertility patients, about 75% or more of the group shows PCOS symptom, causing repeated abortions. Further, it has been reported that an increase in frequency of solenoma and breast cancer due to continuous stimulation of estrogen, arteriosclerosis due to lipid change by raised female hormone level, and frequency of diabetes mellitus is high for accompanied hyperinsulinism. Also, PCOS is the most frequent cause for anovulation, and accompanied by fatal complications, indicating a clinical importance. PCOS is being diagnosed based on clinical symptoms, blood test or ultrasonography, and can be  diagnosed by detecting abnormal menses such as oligomenorrhea or amenorrhea, clinical symptoms such as hypertrichosis, obesity, infertility and acne, and unbalance of female hormone level. PCOS is a disease which causes a variety of complications such as infertility when not suitably treated by screening in adolescent period.
Pertti et al. reported (Fertility & Sterility. 1996. 65(3):517-522) that as a method for differentiating polycystic ovary syndrome (PCOS) from normal women, comparative analysis on the level of luteinizing hormone (LH), follicle stimulating hormone (FSH), and androstenedio shows sensitivity of 98%, specificity of 93%. Tayler et al. reported that 75% or more of PCOS showed increased LH level, and 94% thereof showed a raise in the ratio of LH/FSH (J. Clinical Endocrinology & Metabolism. 1997. 82(7):2248-2256). As such, in a diagnosis of PCOS, the ratio between LH and FSH is used as significant result. However, such a method where the ratio of LH and FSH is determined after analyzing individual level of FSH and LH respectively by analytical instrument, is not suitable as a first screening method, thus at present, no analytical method is available by which PCOS-risk group can be early screened by one step test.
Korean Patent No. 0403871 discloses a device for diagnosing normal pregnancy and ectopic pregnancy, and manufacturing methods thereof, which device analyzes intact human chorionic gonadotrophin (hereafter, referred to as hCG) and modified hCG by using a combination of monoclonal antibodies. According to said disclosure, ectopic pregnancy is diagnosed based on measuring the ratio of intact hCG and modified hCG, where said similar structural proteins in test sample are assayed by use of different set of antibodies respectively. However, said Korean patent No. 0403871 has defects that an instant measurement of the ratio of the similar structural proteins is not possible because the ratio can only be determined after conducting individual analysis on the level of each protein by use of a different set of monoclonal antibodies.
Also, U.S. Pat. No. 5,786,220 discloses a manufacturing method for one-step diagnostic reagent which differentiates normal pregnancy and abnormal pregnancy, and describes that simultaneous determination of the levels of progesterone and hCG in female fluid enables diagnosis of normal pregnancy,  abortion, ectopic pregnancy and cancer etc. That is, according to the disclosure, when the level of progesterone in blood is not more than 25 ng/ml and at the same time, that of hCG is in a range of 25-2,500 mlU/ml, diagnosed as spontaneous abortion and ectopic pregnancy, and when the level of progesterone in blood is 25 or more ng/ml and that of hCG is 2,500 or more mlU/ml, diagnosed as normal encyesis.
International Laid-open Publication No. 0070094 discloses a diagnostic method for trophoblast or non-trophoblast malignancy using antibody specifically reacting with early pregnancy associated molecular isoform (EPMI). According to the disclosure in said International Laid-open Publication No. 0070094, the levels of early pregnancy associated molecular isoform and intact form hCG in test sample are measured by reaction of early pregnancy associated molecular isoform and antibody, and the ratio between them is calculated, thereby to differentiate, normal pregnancy and abortion, trophoblastic disease and non-trophoblastic disease in early pregnancy.
Known methods as described above, that is, method of measuring doubling time of hCG level (during gestational ages of 5-9 weeks, blood level of hCG doubles every 1.4-2 days) and method of diagnosing ectopic pregnancy by measuring the level of free β-hCG in pregnant woman's fluid, are defective for necessity of repeated blood-collecting. Also a diagnostic principle of said methods differs from that of the present invention in that along with the measurement of hCG level, the level of progesterone should be also determined, and said methods include problems that though normal pregnancy and abnormal pregnancy can be differentiated, ectopic pregnancy which is very dangerous disease to pregnant women and spontaneous abortion are difficult to be effectively screened at an early stage.
On the other hand, prostate specific antigen (hereinafter, referred to as PSA) is a protein secreted from normal epithelial cell or cancer cell of prostate, and an enzyme which under normal situation acts to dissolve aggregated semen at the time of ejaculation. Though PSA is normally found only in prostate, when the normal structure of prostate is being destroyed due to tumor or infection, it is to be found even in blood stream. Further, in case of infection such as prostatitis or when prostate is enlarged for prostatomegaly yet not tumor, increased blood PSA level is observed. According to American Cancer  Society, prostatic carcinoma has been reported to be the cancer most frequently diagnosed in men, and in 1999 about 179,300 cases of prostatic carcinoma were diagnosed and prostatic carcinoma shows a high death rate (37,000 persons) in men except lung cancer. Since PSA was lately found to have several molecular structures in blood serum, PSA test has been rapidly developed. The majority of PSA in blood serum binds with a variety of protease inhibitors, i.e. ACT (Alpha 1 antichymotrypsin) and AMT (alpha-2-macroglobulin). PSA-ACT conjugate is a major form in serum and called conjugate PSA. Another form of PSA is also found, enzymatically inactivated non-conjugate form with nothing bound. This is called free PSA. PSA test is performed via analysis of free PSA and total PSA, it was widely known that determining the ratio of free PSA can raise sensitivity and specificity of prostatic carcinoma. Measurement of free PSA is meaningful in that prostatic carcinoma and benign disease can be distinguished when prostatic carcinoma is not suspected on rectal examination yet the level of total PSA is 4.0-10.0 ng/ml. Also the lower the level of free PSA is, the higher the probability of prostatic carcinoma is, and thus measurement of free PSA can reduce unnecessary biopsy by 20-40%, and enables accurate diagnosis of tumor by specificity of 90% or more. Further Klaus (Clinical Chemistry, 2000. 46(1):47-54) et al. reported that tests on the ratio of PSA-ACT and free PSA are also used as marker to differentiate prostatic carcinoma and benign disease.
As described above, the known method performed two tests respectively and determined the ratio of similar structural proteins by comparing the analytical results, thus it has problem that standardization is impossible for a large variation of absolute value depending on the feature of diagnostic device.